TWI225490B - Bioactive whey protein hydrolysate - Google Patents
Bioactive whey protein hydrolysate Download PDFInfo
- Publication number
- TWI225490B TWI225490B TW088110663A TW88110663A TWI225490B TW I225490 B TWI225490 B TW I225490B TW 088110663 A TW088110663 A TW 088110663A TW 88110663 A TW88110663 A TW 88110663A TW I225490 B TWI225490 B TW I225490B
- Authority
- TW
- Taiwan
- Prior art keywords
- patent application
- scope
- item
- hydrolysis
- enzyme
- Prior art date
Links
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J3/00—Working-up of proteins for foodstuffs
- A23J3/30—Working-up of proteins for foodstuffs by hydrolysis
- A23J3/32—Working-up of proteins for foodstuffs by hydrolysis using chemical agents
- A23J3/34—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes
- A23J3/341—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes of animal proteins
- A23J3/343—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes of animal proteins of dairy proteins
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- General Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Nutrition Science (AREA)
- Biochemistry (AREA)
- Zoology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Polymers & Plastics (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Peptides Or Proteins (AREA)
Abstract
Description
技術領域: 本發明係關於一種萝-不呈^ 1 Γ ——〜η 肽類之到生恭ώ u 不,、古味且含有具丨生物if性丨之艇 肢類之礼α蛋白巧水解物的方法。此方氺Ϊ 一古玄 η ^ ^21 V- Ah 1, 凡万法之產物有呵 >為化率 及良好的\ g月色性。此箄產物可处 曰ϋ—m ι 寺座物了犯具有溫和或些微的甜味, 窠醣,I Α ^未此寻礼4蛋白水解物可視需要含有 來源。 、铖此性艮口口中之生物活性胜肽類之有致 背景技藝 許多食品成份及食物已經是 乳蛋白,酪蛋白及乳清蛋白等 在健康照護的許多領域中, 水解蛋白質食物之優勢。舉例 白質較不具過敏原性。其亦能 吸收。含有水解蛋白質之食物 患者之攝食也有助益。 由水解蛋白質來源,如:牛 而製成。 水解蛋白質食物具有超越非 而言,已知經酵素水解之蛋 車父全蛋白更快速地被消化及 對醫院病患如具消化性疾病Technical Field: The present invention relates to a kind of glutinous-non-presenting ^ 1 Γ ~~ η peptides. No, antique, and containing 丨 bio-if 丨 boat polite gift α protein hydrolysate Methods. This side is an ancient mystery η ^ ^ 21 V- Ah 1, the product of Fanwanfa has a chemical conversion rate and good moonlight properties. This product can be described as "ϋ"-m ι temple seat has a mild or slightly sweet taste, caramel, I ^ ^ ^ this protein 4 hydrolysate may contain the source as required. This is because of the biologically active peptides in the mouth. Background technology Many food ingredients and foods are already milk protein, casein, and whey protein. In many areas of health care, hydrolyzed protein foods have the advantage. Examples White matter is less allergenic. It can also be absorbed. Foods containing hydrolyzed protein Food intake by patients also helps. Made from hydrolyzed protein sources such as cattle. Hydrolyzed protein food has more than that. For example, the enzyme-hydrolyzed egg is known to be digested more quickly by whole car protein.
已知將乳清蛋白及酪蛋白之水解可釋出呈現數種生理效 用的生物活性胜肽類(Maub〇is等人,1991,歐洲專利第 4755 0 6 )號。許多刊物述及此等具生物活性之胜肽類,例 如·經由酵素處理之牛血清沒—乳球蛋白及乳清蛋白濃縮 物而得到之具有抗高血壓性之ACE抑制胜肽(MuUally等 人,1 997 )。ACE抑制胜肽亦發現存在於αg及召—酪蛋白之 水解物中(日本專利第4282400號;Nakamura等人,1 994, Yamamoto 等人,1994)。 歐洲專利第4745506號揭示乳清中之牛乳乳鐵蛋白水解It is known that hydrolysis of whey protein and casein can release bioactive peptides exhibiting several physiological effects (Maubois et al., 1991, European Patent No. 4755506). Many publications mention these biologically active peptides, such as the anti-hypertensive ACE inhibitory peptides obtained from enzyme-treated bovine serum-lactoglobulin and whey protein concentrates (MuUally et al. , 1 997). ACE-inhibitory peptides are also found in hydrolysates of αg and tau-casein (Japanese Patent No. 4282400; Nakamura et al., 1 994, Yamamoto et al., 1994). European Patent No. 4745506 discloses hydrolysis of bovine milk lactoferrin in whey
第4頁 五 發明說明(2) 可以釋出乳鐵蛋白冬, ’舄’、香港腳’眼部感染,;l :當;:治療人類或動物之腹 然而,已知大部份 、決病之抗微生物劑。The fifth page of the invention description (2) can release lactoferrin winter, '舄', Hong Kong feet 'eye infections; l: when ;: treatment of the abdomen of humans or animals Antimicrobial agent.
Si產:之水解解^其是含有乳清及 =口以蛋白水解物⑵=: f ^白貝水解之領域中,通常使一 =來控制或去除蛋白f水解物:種或同時使用兩種 味性。 未,以增加產品的美 已知廣泛地水解蛋白質基質可 味(歐洲專利第0 65663 ’1 1 7047號,美礼蛋白水解物之苦 號)。此法可較簡易且較便宜地生產‘”第39·0 而,廣泛地水解會輪短所有胜狀 -古味之產品。然 具有生物活性之胜肽。廣泛地水:質=::得到之 得到之胜肽的機能性及生物活性。此外貝欲 質基質之溫和味道仍較差相=來蛋白 而f ’苦味只部份被去除(Roy 1992及1 9 9 7 )。t水解物 味【音種:t控!1蛋白質水解物苦味的方法是利用去苦 者’、 ,、是彼等來自米麴菌(Aspergillus 〇ryzae) ,蛋白質水解所產生之”苦味”認為係肇因於大疏水性,,苦 "f 胜肽之存在。去苦味酵素可選擇性地水解存在於蛋白 質水解物中之苦味胜肽。熟習此技術之工作者可藉由明智Produced by Si: Hydrolyzed ^ It contains whey and = protein hydrolysate ⑵ =: f ^ in the field of white shell hydrolysate, usually use a = to control or remove protein f hydrolysate: species or use both Flavor. In order to increase the beauty of the product, it is known to extensively hydrolyze the taste of the protein matrix (European Patent No. 0 65663'1 1 7047, Merlot protein hydrolysate). This method can be simpler and cheaper to produce "" Article 39 ·. Extensive hydrolysis will shorten all the products of the vicissitude-antique taste. However, it has biologically active peptides. Extensive water: quality = :: obtained The functional and biological activity of the obtained peptide. In addition, the mild taste of the shellfish matrix is still poor. Phase = protein and f 'bitterness is only partially removed (Roy 1992 and 197 7). Species: t control! 1 The method for bitterness of protein hydrolysates is to use bitters, ',, and they are from Aspergillus 〇ryzae. The "bitterness" produced by protein hydrolysis is thought to be caused by large hydrophobicity. The existence of bitter peptides. Debittering enzymes can selectively hydrolyze bitter peptides present in protein hydrolysates. Workers familiar with this technology can use wise
第5頁 1225490 發明說明(3) =及處理條件,將牛乳蛋白水解物中之苦 _ :然";留下所欲得到具生物活性之胜 且斗仏X ^ 使用去古味酵素使得製程更加昂貴,且一些 得。Η = = Τ來或被成功地獲 酸至終產物之趨向,結果具有釋出游離胺基 味或4味。UQy 1 992及1 997 ) 7 生令人*悅之肉湯 各式各樣的去除蛋白水解 額外製造步驟及增加势送=古2方法均導致終產物之 酸終產物變成超出平;?成本…卜,由於產生游離胺基 倘若能開發出一種水組I ^I 物活性之胜肽類耧ψ B蛋白貝之方法,使所欲得到具生 成,可限制苦味胜狀及游離胺基酸生 大之優Ϊ 質之溫和味叇得以保冑,則將具有莫 一些具生物活性之胜肽類 ^ 質水解過程中相對地較為安定且非常二血壓胜肽-在蛋白 之水解過程中釋放出來(如圖…』):易於牛乳蛋白基質 牛乳蛋白水解物的苦味可藉由添加 牛乳蛋白基質中之天然糖類( ^員或水解已存在於 (Β⑽amlen,1 98 9 )。例如 ^主而加以改善 ^ 一半乳糖甘酶及乳糖鎢將乳J = = =清若藉 其更具美味性(法國專利第2309 1 54號;m匕,可使 4358464號;日本專利第㈣咐^號)。關專利第 為了使含有具生物活性之胜肽類的蛋白質水解產 1225490Page 5 1225490 Description of the invention (3) = and processing conditions, the bitterness in the milk protein hydrolysate _: Ran " leave the desired biologically active victory and 仏 仏 X ^ use of anti-palatase enzyme to make the process more Expensive and some too. Η = = T tends to be or is successful in obtaining the acid to the final product, with the result that it has a free amine or 4 flavor. UQy 1 992 and 1 997) 7 Delightful broths of all kinds Remove proteolysis Extra manufacturing steps and increase potential = ancient 2 methods have all caused the acid end products of the final products to become more than flat; Cost ... By, because the production of free amine groups can be developed if a peptide of the water group I ^ I activity is active 耧 ψ B protein shell, so that the desired production can be achieved, the bitter taste and free amino acids can be limited Shengda Zhiyou, the mild miso of quality can be preserved, then some peptides with biological activity will be relatively stable ^ during the hydrolysis of the protein is relatively stable and very ditensive peptides-released during the hydrolysis of the protein (As shown in the picture ...): The bitter taste of cow milk protein hydrolysate which is susceptible to the milk protein matrix can be improved by adding natural carbohydrates in the cow milk protein matrix (^ members or hydrolysis is already present (B⑽amlen, 1 98 9). ^ Galactosidase and Tungsten Lactose J = = = Qing Ruo will be more delicious (French Patent No. 2309 1 54; m dagger can make No. 4358464; Japanese Patent No. ^). Related patents The first is to hydrolyze proteins containing bioactive peptides to produce 1225490.
五、發明說明(4) 高度的風味可接受性,有必要精確地控制水解過程以避免 苦味生成。 一種常用來終止水解反應的方法是藉由將酵素去活性 化,通常在高溫藉熱去活化,於9 0〜丨0 〇 〇c以上維持/一段時 間。然而,此方法無法用來終止乳清蛋白之水解,'因^ ^ 在於混合物中之完全未被水解的乳清蛋白會而變性且沉 殿,使得終產物之溶解度較差且其用來作為食品成份2接 受度亦較牴。 倘若有一種製造水解乳清蛋白質之方法,可經控制使得 直接生成含有生物活性胜狀類以供添加於機能性食品之水 解物,其不具苦味且酵素之去活性化步驟不會破壞Z產物 中所含之完整蛋白質,將會是有利的。 本兔明之一目的即疋提出一些有法,以期逵到此蓉兩 求,或者,至少提供大眾一個有用的選擇?達到此寺而 發明概述 因此,本發明可廣泛地述及製備含有生物活性胜肽之乳 清蛋白水解物的方法,其包括以下步驟: (i )以一或多種可水解乳清蛋白之酵素處理乳清蛋白基 質,以生產一種含有生物活性胜肽之乳清蛋白水解物;及 (i i)於苦味風味實質生成之前終止水解反應。 &較佳地將水解作用在最多只造成部份乳清蛋白變性且仍 能維持或改善此水解物之官能性的反應條件下終止。 更佳地’水解作用能在部份乳清蛋白變盖 官能性的反應條件下被終止。 。一5. Description of the invention (4) High flavor acceptability, it is necessary to precisely control the hydrolysis process to avoid bitterness. A commonly used method to stop the hydrolysis reaction is to deactivate the enzyme, usually by deactivating it at high temperature, and maintaining it for more than 90 ° C to 0 ° C for a period of time. However, this method cannot be used to stop the hydrolysis of whey protein. 'Because the completely unhydrolyzed whey protein in the mixture will denature and sink, making the final product poorly soluble and used as a food ingredient 2 Acceptance is also relatively low. If there is a method for producing hydrolyzed whey protein, it can be controlled to directly generate a hydrolysate containing biologically active species for addition to functional foods, which does not have a bitter taste and the enzyme deactivation step does not destroy the Z product. The intact protein it contains will be beneficial. One of the goals of this rabbit is to propose some ways to achieve this, or at least provide a useful option for the public? Summary of the invention upon reaching this temple Therefore, the present invention can broadly describe a method for preparing a whey protein hydrolysate containing a biologically active peptide, which includes the following steps: (i) treatment with one or more enzymes that hydrolyze whey protein A whey protein matrix to produce a whey protein hydrolysate containing a bioactive peptide; and (ii) stopping the hydrolysis reaction before the bitter flavor is substantially generated. & The hydrolysis is preferably terminated under reaction conditions which at most cause denaturation of part of the whey protein and still maintain or improve the functionality of this hydrolysate. More preferably, the 'hydrolysis can be terminated under a reaction condition where part of the whey protein becomes functional. . One
第7頁 1225490 五、發明說明(5) 較佳地,用以水解乳清蛋白的酵素可選自於下列所組 之群:蛋白酶P6,蛋白酶A,蛋白酶M,胜肽酶,中性 酶’有效酶(Validase)及AFP 200.0 (均如本文所定 水解之終止以熱處理較佳,處理時間較佳以i至1〇秒, 在溫度約8 5 °C至1 0 0 °C。 水解終止前之基質水解程度以至多丨〇%較佳。 水解程度以約3%至約5%更佳。 基質以含乳糖達50%重量比者較佳。 或基質中含乳糖達3 0 %者亦可。 、基質以經由乳糖酶及/或点-半乳糖苷酶處理者較佳, i者ΐ於乳清蛋白水解前或水解期間,將乳糖水解成半朝 糖及葡萄糖並合成半乳募糖。 本=明之另一較佳具體實施㈣是一種乳清蛋白水解 AFFΓ或多種生物活性胜肽,選自下列所組成之群: VTSnv ^ ’ IUEKH,UVTQ,MKG,LDIQK,VF,ALP_, vts^av ,lhlplp ,LVYPFPGPIPNSLPQNIpp及LFRQ 。 之理nn:藉由熟習此等技術的人對某種特定酵辛 之=解’而於適合該特定酵素之條件下進行。 阵素 :用於乳清蛋白濃度為:固形物5〜5〇% 總固形广為"酵素二基質比介於。.。崎。(重量/ 重量比)者^佳比)其中以介於〇.〇1%至1%(重量/總固形物 性蛋白酶處理蛋白質基質,可於ρΗ 25 進仃水解,並以ρΗ 3.0〜5.〇之間較佳。 之間Page 7 1225490 V. Description of the invention (5) Preferably, the enzyme used to hydrolyze whey protein may be selected from the group consisting of protease P6, protease A, protease M, peptidase, neutral enzyme ' Validase and AFP 200.0 (both as stated herein, the termination of hydrolysis is better by heat treatment, the treatment time is preferably from 1 to 10 seconds, at a temperature of about 8 5 ° C to 100 ° C. Before the termination of hydrolysis The degree of matrix hydrolysis is preferably at most 10%. The degree of hydrolysis is preferably about 3% to about 5%. The matrix is preferably 50% by weight with lactose or the matrix contains 30% lactose. The substrate is preferably treated with lactase and / or dot-galactosidase, and those who have hydrolyzed lactose to hemichalcose and glucose and synthesize galactose before or during the hydrolysis of whey protein. = Another preferred embodiment of Ming is a whey protein hydrolyzed AFFΓ or multiple biologically active peptides selected from the group consisting of: VTSnv ^ 'IUEKH, UVTQ, MKG, LDIQK, VF, ALP_, vts ^ av, lhlplp, LVYPFPGPIPNSLPQNIpp, and LFRQ. nn: People who are familiar with these technologies The specific enzyme is the solution. It is carried out under the conditions suitable for the specific enzyme. Matrix: used for whey protein concentration: solids 5 ~ 50% The total solids is widely known as the enzyme two matrix ratio .. . Saki. (Weight / weight ratio) is better.) Among them, the protein matrix is treated with 0.001% to 1% (weight / total solid physical protease), which can be hydrolyzed at ρ 25 and ρ 3.0 3.0 ~ It is better between 5.
第8頁 1225490 五、發明說明(6) 若以中性蛋白酶處理蛋白質基質,可於ρΗ 3 · 5〜9 . 〇之間進 行水解,並以pH 6· 0〜8· 0之間較佳。 若以鹼性蛋白酶處理蛋白質基質,可於pH 5· 0〜10. 〇之 間進行水解,並以p Η 6 · 0〜8 · 0之間較佳。 蛋白質水解可於溫度範圍20-65 °C下進行,並以50-60 °C 較佳。 乳糖水解之進行可進行於乳清蛋白水解之先前階段,同 時ί ^後。用來水解乳糖的酵素可為乳糖酶及/或冷—半乳 ^ It 且其可遥自酵母或真菌來源,如:Klyvermyces Saccharomyces lactis, Saccharomyces fragillls,如米麴菌,黑麴菌,如 des)及Novolact(Novc) Nordisk)。乳糖水解於孰習 此技術之人員所知之條件下進行+ …、 一種終止水解方、、土 一 蛋白水解酵素式此::貫施例乃藉由使一或多種乳清 化來達成,首弁获2先刖加入之乳糖水解酵素)之去活性 失活或較不具活=將反應混合物之酸鹼值改變至使酵素 合物加熱至幸交、^ 4之酸鹼值,及/或用熱交換器將反應混 之完整乳清蛋白變里度,以使酵素變性但卻不使基質中 70 °c,較佳為65。7。。酵素失活的適當溫度範圍為55- 根據一選擇,與所 燥之步驟使酵素 用之酵素有關,亦可使用蒸發或乾 根據另一選擇,素混合物失活。 整酸鹼值而去活化酵素或酵素混合物可以先前調整或不調Page 8 1225490 V. Description of the invention (6) If the protein matrix is treated with a neutral protease, it can be hydrolyzed between ρΗ 3 · 5 ~ 9. 〇, and preferably between pH 6 · 0 ~ 8 · 0. If the protein matrix is treated with alkaline protease, it can be hydrolyzed between pH 5.0 · 10 ~ 1.0, and preferably between pΗ6 · 0 ~ 8 · 0. Proteolysis can be performed at a temperature range of 20-65 ° C, and preferably 50-60 ° C. The lactose hydrolysis can be carried out at a previous stage of whey protein hydrolysis, and at the same time. The enzymes used to hydrolyze lactose can be lactase and / or cold-galactose ^ It and it can be remote from yeast or fungal sources, such as: Klyvermyces Saccharomyces lactis, Saccharomyces fragillls, such as Oryzae, black fungus, such as des) And Novolact (Novc) Nordisk). Lactose hydrolysis is carried out under conditions known to those skilled in the art +, a method for terminating hydrolysis, and a protein-hydrolyzing enzyme formula: This example is achieved by making one or more whey, first (Get 2 lactose hydrolyzed enzymes added before) Deactivation inactivation or less active = change the pH value of the reaction mixture to heat the enzyme compound to Xingjiao, pH value of 4 and / or use The heat exchanger mixes the reaction with intact whey protein to denature the enzyme without denying 70 ° C in the matrix, preferably 65.7. . The appropriate temperature range for enzyme inactivation is 55- depending on the choice, depending on the enzyme used in the drying step, or by evaporation or drying. According to another choice, the inactivation of the vegetarian mixture. Adjust pH or deactivate enzymes or enzyme mixtures can be adjusted previously or not
第9頁 1225490 五、發明說明(7) —或者,用以選擇性水解乳清蛋白的一種或 ::二ΐίΓϊ’如:RoehmEupergit,鹿角Ϊ“ 粒,成丁聚醣顆粒或任何其它合適的物質上=:膠顆 器。 膜上,或在巾空纖維管反應 此素可於水解作用進行前,A 方式固定於適當的鈍性支持物上 ^鏈、之 濾膜加以分離。 1γ κ解作用後以微過 或者,酵素可於水解完成時,立 1 〇- 5 0 0仟道耳頓(KDa)之超過滹/ :/慮過分子量 來。 愿朕自水解混合物中分離出 在水解及視需要使酵素失活或移 物可視需要在條件下進行逆滲透萨此自j,此等水解 類及水份。如此便可回收含有乳清‘白除鹽 水解物中還會含有葡萄糖,=;;將:;唐加,水解,則 含有具生物活性之胜肽類的乳清 3水解< I糖。 使用可遽過分子量5_200仟冑耳頓^^水^物亦可視需要 較佳為10-50仟道耳頓之。具生物活=分離出來, 肽類及,視需要地,被水解的乳糖丨肽類,其他胜 收。 胖扪礼搪均可於過濾液中被回 根據另一選擇,離子交換樹脂或 互作用層析法或此等方法之組合亦;於=或疏水*** 而得到較強化型式的具生物活性部份。;7解物中回收Page 9 1225490 V. Description of the invention (7)-Or, one or selectively used to selectively hydrolyze whey protein, such as: "Roehm Eupergit, Staghorn", granules, into butanose particles or any other suitable substance Top =: gel particles. On the membrane, or before the reaction of the hollow fiber tube, the element can be fixed on the appropriate blunt support ^ chain, and the membrane is separated before the hydrolysis. 1γ κ solution After that, the enzyme may be passed by when the hydrolysis is completed, and the molecular weight will be more than 10/500 (Da) (KDa), and the molecular weight will be taken into consideration. Need to make the enzyme inactivated or move the material under the conditions of reverse osmosis, such as j, such hydrolysis and water. So you can recover whey 'white desalted hydrolysate will also contain glucose, =; Will be: Tanga, hydrolyzed, whey 3 containing bioactive peptides hydrolyzed < I sugar. The use of a molecular weight of 5 to 200 仟 胄 仟 胄 ^ ^ water ^ may also be better if necessary 10 -50 仟 Doorton. With biological activity = isolated, peptides and, if necessary Hydrolyzed lactose 丨 peptides, others win. Fat can be returned in the filtration solution. According to another option, ion exchange resin or interaction chromatography or a combination of these methods is also available; in = or hydrophobic Sexual intercourse to obtain a stronger type of biologically active part .; 7 solution recovered
O:\58\58905.ptd 第10頁 1225490 五 發明說明(8) __ 以乳糖…-半乳糖苔酶水解 : +礼券糖已知其可促進腸内有益菌之生 斤產生之 水解物之生物活性特性。 、 匕亦可提高 進-步水解乳糖水解物對乳糖 者而έ ,是很有用的食品添加物。 了足之4費 本發明之水解乳清蛋白產品具有—或 • t =壓性之升壓素轉換酵素抑制劑⑽」ΪΓ •雙歧桿菌生長促進活性 11 •不黏,不具苦味 • 具有悅人的至輕微的甜味 •良好的官能性 二!:包括上述之說明及後述之實例在内。 圖式間要說明 一 現在將對本發明之附圖進行說明,其中. 圖1是一以苦味及生物活性為縱軸/以水 =圖。根據本發明而於苦味胜肽類生成之前庐王二為橫軸 活性之胜肽類並具有可接受性 二::有具生 於線\及線x2之間。 禾之產的機會窗"介 表示四組以不同飼料餵食8週之 期血壓變化圖。 大白以的收縮 银Γ八為么曰以餵食商品化大白鼠飼料為控制組,相料 均值分析圖。 的幻枓之大白鼠的最小平方 又义口口 . 1225490 五、發明說明(9) 如刖文所述,本發明提供一 之乳清蛋白水解物之方法,苴衣^ 3生物活性胜肽類 π避免在水解期間產生;;良解如作用=高度^制下 =。水解作用如圖i所示,於 =,及 即於大量苦味生成前即終止 中被終止’ 最大量之生物活性胜肽類:供:2m性及 於圖1中之X軸。介於赴v sa K解私度定Ϊ地表示 素而變動。最適水解條件即\曰使的苦機會窗將^ 生物活性達最高量。 未在了接文之範圍内而 酵ΐίΓ=;!Τ圭具體實 <列,水解乳清蛋白所用的 坪系你&自下列所組成之群:白, ^ 醜’胜肽酶’”生酶,有效酶及AFP2_(=本= 疋,且藉由南溫短時之熱處理X約85- 1 0 0。〇處理卜! 〇 秒)終止水解作用。於此應用中可喜地發現上述酵素(1 生產含不少量生物活性胜肽類之乳清蛋白水解物(2)可於 短時之熱處理下失活,處理過程中只有部分乳清蛋白 吏、’且令人驚喜地改善乳清蛋白之官能性,包括提供具 有滑潤口感(因其顆粒相對較小)且外觀相當潔白。八” 現在將本發明以下列實例舉例說明之 實例1 蛋白質含量為80%之甜乳清蛋白濃縮物之1〇%溶液 ULACENTM3 92,2公升),於50 °C下,以由枯草桿菌生產之 商品化酵素:中性酶(N〇v〇 N〇rdisk,丹麥)水解,水解O: \ 58 \ 58905.ptd Page 10 1225490 Five descriptions of the invention (8) __ Hydrolysis with lactose ...- galactosidase: + Gift coupon sugar is known to promote the hydrolysate production of beneficial bacteria in the intestine Active properties. It can also improve the further hydrolysis of lactose hydrolysate, and it is a very useful food additive. The full cost of the hydrolyzed whey protein product of the present invention is-or t = compressive vasopressin-converting enzyme inhibitor ⑽ "ΪΓ • bifidobacterium growth promotion activity 11 • non-sticky, no bitter taste • pleasant To slight sweetness • Good functionality two! : Including the above description and examples described later. The drawings will be explained in the following. The drawings of the present invention will now be described. Among them, FIG. 1 is a graph with bitterness and biological activity as the vertical axis / water = graph. According to the present invention, before the generation of bitter peptides, Luwang II is a peptide with horizontal axis activity and is acceptable. 2: It is born between line \ and line x2. The window of opportunity of He Zhichan said that the blood pressure changes of the four groups were fed with different feeds for 8 weeks. The shrinkage of Dabai is based on the analysis of the mean value of the average data of the feedstock of commercial rats as the control group. The least square of the phantom of the rat is 1225490 V. Description of the invention (9) As described in the text, the present invention provides a method of hydrolysate of whey protein, 苴 衣 ^ 3 biologically active peptides π avoids production during hydrolysis; good solutions such as action = height ^ system under =. The hydrolysis is shown in Figure i, at =, and is terminated before a large amount of bitterness is generated. The maximum amount of biologically active peptides: for: 2m, and on the X axis in FIG. It varies depending on the expression of v sa K. The most suitable hydrolysis conditions are the window of opportunity for suffering and the highest biological activity. It is not within the scope of the text to leave the ΐΓ = ;! Τ 圭 concrete implementation list, which is used to hydrolyze whey protein. The group consists of the following: white, ^ ugly 'peptidase' Enzymes, effective enzymes and AFP2_ (= this = 疋, and by the short-term heat treatment of South temperature X about 85-1 0 0. 0 treatment Bu! 0 seconds) to terminate the hydrolysis. In this application, the above enzyme can be found with joy (1 Production of whey protein hydrolysates containing not a small amount of bioactive peptides (2) can be inactivated by short-term heat treatment, only part of the whey protein is processed, and the whey protein is surprisingly improved Functionality, including providing a smooth mouthfeel (because of its relatively small particles) and a fairly white appearance. Eight "Example 1 of the present invention is exemplified by the following examples: 1 of a sweet whey protein concentrate with a protein content of 80% 〇% solution ULACENTM3 92, 2 liters), at 50 ° C, hydrolyzed with a commercial enzyme produced by Bacillus subtilis: neutral enzyme (NovvNordisk, Denmark), hydrolyzed
O:\58\58905.ptd 第12頁 1225490 五、發明說明(10) 水解物之酸鹼值調為5· 〇並於65 t加熱3〇分鐘以使酵素失 =°續將水解物(水解度為4· 5%)加以噴霧乾燥並測試其升 壓素轉換酵素抑制劑(ACE- I)活性及風味。乾燥產品之 ACE-Ι 活性根據Dw Cushman 及HS Cheung (1971)之方法, 以FAPGG(產品305-1〇 , Sigma Chemical Corporation ,O: \ 58 \ 58905.ptd Page 12 1225490 V. Description of the invention (10) The pH value of the hydrolysate is adjusted to 5.0 and heated at 65 t for 30 minutes to lose the enzyme = ° (4.5%), spray-dried and tested for its vasopressin-converting enzyme inhibitor (ACE-I) activity and flavor. The ACE-I activity of the dried product was determined according to the method of Dw Cushman and HS Cheung (1971) with FAPGG (product 305-1, Sigma Chemical Corporation,
St· Louis ’ USA)為基質測定之。其活性以使ACE_i活性降 低一半所需之物質量(公克/公升)表示之。此水解物之IC5q ACE-I活性為〇· 44公克/公升且經味覺試驗而測得之風味接 受度分數相當高。 實例2 ^ ALACENTM 421乳清蛋白濃縮物之50%溶液(蛋白質含量 56% 公升以由Kluveromyces lactis生產之商品化酵 素 乳糖酶(Lactozyme 3000L,Novo Nordisk)於酵素 / 基 貝比為0 · 3 /,5 〇 C下,處理2小時。再將此溶液以中性酶 · 於酵素/基質比為〇· 3%,50 °C下,水解1小時。之後,將混 合物稀釋五倍’藉超高溫瞬間加熱處理(95 °C,5秒)使活 -性^素失活。續將該水解物喷霧乾燥。乾燥粉末(水解度 2· 8%)並不含任何活性酵素且具有ACE-Ι活性為2· 18公克/ f升。其風味接受度分數因產品受到低度之甜化而例外地 南。ACE- I測定及風味接受度之評分方法同實例1。 〇 實例3 以與貫例1相似之方法,由ALACENTM 3 9 2製得之5 0 0公升 水解物於酵素失活後,冷卻至1 0 °C。將原始水解物取出一 部伤加以乾燥’剩餘水解物在1 〇 °C以能截切分子量1 0,〇 〇 〇St. Louis' USA) was determined by matrix. Its activity is expressed in terms of the mass (g / litre) required to halve the activity of ACE_i. The IC5q ACE-I activity of this hydrolysate was 0.44 g / liter and the taste acceptance score measured by a taste test was quite high. Example 2 ^ 50% solution of ALACENTM 421 whey protein concentrate (56% liters of protein with a commercial enzyme lactase (Lactozyme 3000L, Novo Nordisk) produced by Kluveromyces lactis at an enzyme / kibe ratio of 0.3 /, Treatment at 50 ° C for 2 hours. This solution was then hydrolyzed with a neutral enzyme · in enzyme / matrix ratio of 0.3% at 50 ° C for 1 hour. After that, the mixture was diluted five times' by ultra-high temperature instant Heat treatment (95 ° C, 5 seconds) deactivates the active-active hormone. The hydrolysate is spray-dried. The dry powder (degree of hydrolysis 2.8%) does not contain any active enzymes and has ACE-1 activity. It is 2.18 grams / f liter. Its flavor acceptance score is exceptionally low due to the low degree of sweetening of the product. The ACE-I measurement and flavor acceptance score method is the same as in Example 1. 〇Example 3 is the same as in Example 1. In a similar way, 500 liters of hydrolysate made from ALACENTM 3 92 was cooled to 10 ° C after the enzyme was deactivated. The original hydrolysate was taken out and wounded, and the remaining hydrolysate was at 10 ° C. In order to cut the molecular weight 10,000
第13頁 1225490 五、發明說明(11) 道耳頓之過渡膜(HFK 131,Koch Membrane Systems, USA)進行超過濾。該水解物(水解度介於3 · 8%至4· 2%之間) 被濃縮至V C F 1 0且濃縮液直接進行乾燥。濾過液以蒸發法 濃縮至2 5 %固形物並加以乾燥。A C Ε - I測定及風味接受度之 評分係如實例1測定。濾過液粉末之ACE- I活性提高(濾過 液粉末之I Cm為0 · 1 5公克/公升)。於超過濾之前取出之原 始水解物,經乾燥後之ACE-Ι活性為〇· 43公克/公升。濃縮 液乾燥粉及噴霧乾燥粉之風味接受度均高。 實例4 得自 ALACEFM 392,ALACE 評 421 及 ALACENTM 392 和乳糖 φ 之混合物之三種不同溶液被製成至多為丨5 %固形物水產生 150升。以由枯草桿菌生產之商品化酵素:中性酶(N〇v〇Page 13 1225490 V. Description of the invention (11) Doulton's transition membrane (HFK 131, Koch Membrane Systems, USA) is used for ultrafiltration. The hydrolysate (degree of hydrolysis between 3.8% and 4.2%) was concentrated to V C F 1 0 and the concentrated solution was directly dried. The filtrate was concentrated by evaporation to 25% solids and dried. The A C E-I measurement and flavor acceptance scores were determined as in Example 1. The ACE-I activity of the filtered powder was increased (the I Cm of the filtered powder was 0 · 15 g / litre). The original hydrolysate taken out before ultrafiltration, and its ACE-1 activity after drying was 0.43 g / liter. Both the concentrated liquid dried powder and spray-dried powder have high flavor acceptance. Example 4 Three different solutions from ALACEFM 392, ALACE Rating 421 and a mixture of ALACENTM 392 and lactose φ were made up to 5% solids water to produce 150 liters. With a commercial enzyme produced by Bacillus subtilis: neutral enzyme (Nov.
Nordisk,丹麥)及由Klyveromyce-s lactis生產之商品化 ‘ 酵素:乳糖酶(Lactozyme 30 0 0L,N0V0 Nordisk)處理溶. 溶液。酵素添加率為中性酶〇· 3%重量比(以蛋白質為基準)· 及Lactozyme 1· 2%(以乳糖酶為基準)。反應在ρΗ 7· 〇土, - 50 C下持續2小時,每〇· 5小時取出35公升樣品,以88處 秒鐘使酵素失活後,進行噴霧乾燥。其活性之測 疋如貫例1,二混合溶液以蛋白質為基準,分別為〇· 4公 克/公升,0· 3 36公克/公升及〇· 432公克/公升。得自 ALACENTM 392之樣品的苦味度與兩個標準樣品相比較。若 將苦味度分為1到10分,且以10分為苦味最強者,則水 〇·5及2小時後之樣品的苦味度值分別為19及2·3,而水 程度較大的兩個標準樣品苦味度值分別為5及7。 _Nordisk, Denmark) and a commercial product produced by Klyveromyce-s lactis ‘Enzyme: Lactozyme (Lactozyme 300 0 0L, NOV0 Nordisk) processing solution. The enzyme addition rate is neutral enzyme 0.3% by weight (based on protein) and Lactozyme 1.2% (based on lactase). The reaction was continued at ρ 下 7 · 〇 soil at -50 C for 2 hours. A 35 liter sample was taken every 0.5 hours, the enzyme was deactivated at 88 seconds, and then spray-dried. The activity was measured as in Example 1. The two mixed solutions were based on protein, which were 0.4 g / L, 0.336 g / L and 0.432 g / L, respectively. The bitterness of the sample obtained from ALACENTM 392 was compared with two standard samples. If the bitterness is divided into 1 to 10 points, and the strongest bitterness is divided into 10 points, the bitterness values of the samples after 0.5 and 2 hours of water are 19 and 2.3, respectively, and the larger two The bitterness values of each standard sample were 5 and 7, respectively. _
1225490 五、發明說明(12) 2小時後取出之ALACENTM 421及ALACENTM 3 92和乳糖之混 合物經之樣品分別具有平均粒徑為3微米或2微米。 ALACENTM 3 92之樣品經2小時水解及如上界定之不活化後具 有平均粒徑為6微米。粒徑較小的樣品具較少砂碟及白粉 感。 經水解之A L A C E NTM 3 9 2 /乳糖混合物混合物之溶解度於該 酸鹼值範圍内為最85%。ALACENTM 3 92及ALACENTM 421樣品 之溶解度大約為70%,而在pH 3· 5下,溶解度稍微下降至 6 5% ° 實例5 得自 ALACENTM 392,ALACENTM 421 及 ALACENTM 392 和乳糖 之混合物之三不同溶液被製成至多為3 〇 %固形物以產生7 5 公升。酵素處理係使用如實例4吃相同條件。每隔半小時 所取出之樣品稀釋至固形物含量為1 5 %。除了以實例4進行 · 熱處理。ACE- I活性測定法同實例1,其活性分別為〇 · 5 6 〇 公克/公升,〇· 440公克/公升及〇· 728公克/公升。 添加0 · 1 %濃度之得自實例4及實例5之樣品至1225490 V. Description of the invention (12) Samples taken from the mixture of ALACENTM 421 and ALACENTM 3 92 and lactose taken out after 2 hours have an average particle size of 3 microns or 2 microns, respectively. Samples of ALACENTM 3 92 had an average particle size of 6 microns after 2 hours of hydrolysis and inactivation as defined above. Samples with a smaller particle size have less sand and white powder. The solubility of the hydrolyzed A L A C E NTM 3 9 2 / lactose mixture mixture is at most 85% within this pH range. The solubility of the samples ALACENTM 3 92 and ALACENTM 421 is about 70%, and the solubility drops slightly to 6 5% at pH 3.5. Example 5 Three different solutions from ALACENTM 392, ALACENTM 421 and a mixture of ALACENTM 392 and lactose It is made up to 30% solids to produce 75 liters. The enzyme treatment system used the same conditions as in Example 4. Samples taken every half an hour are diluted to a solids content of 15%. Except in Example 4, the heat treatment was performed. The ACE-I activity was measured in the same manner as in Example 1. Its activity was 0.556 g / L, 0.444 g / L, and 0.728 g / L, respectively. Add samples from Example 4 and Example 5 at a concentration of 0.1% to
Bifidobacterium lactis之標準培養基中及導致菌體生 長較快且該菌之最終菌體量較不添加補充劑之控制組為 高。 ♦ AUCENTM 39 2,ALACENTM 421 及 ALACENTM 392 和乳糖混合 物彼等二種之水解樣品之募醣(參醣或以上)含量分別為 〇· 2%,2· 1% 及7. 〇%。 實例6The standard medium of Bifidobacterium lactis and the growth of bacterial cells were faster and the final bacterial volume of the bacteria was higher than that of the control group without supplements. ♦ The AUCENTM 39 2, ALACENTM 421 and ALACENTM 392 and lactose mixtures of the two hydrolyzed samples had sugar content (ginseng sugar or above) of 0.2%, 2.1%, and 7.0%, respectively. Example 6
第15頁 1225490 五、發明說明(13) 以實例5中所製備之水解物粉末 ,, 甘、夫a旦几田 冬作為酸礼路(yoghurt)之 添加物:其:加罝佔隶終酸乳各之2 5%及5%,結果與控制 組相比較,,、滑潤性增加且質地酶亦有改盖。 實例7 ° 以實例5中所製備之水解物扒古从^ … w初杨末作為密斯里條食譜 (muesli bar recipe)之尽白暂水、Ε + 〈贫白貝來源,添加率為重量比6% 及1 2 %。所有的品嚐者對皮% 7欠 士 曰水解物條的喜好度均 WPC控制組。其中以添加由實例 ' ^ ^ w 々街a例b中所製備之經水解之 ALACENTM 42 1 及ALACENTM ^ 她 A , 和礼糖之混合物者有最好的結 果。 實例8 =貝例5^中所製備之水解物粉末作為代餐概念樣品之組 刀。於礼糖及蛋白質中被水解7的aucentm 42ι以佔重量 力:率添加至全乳粉,麥芽糊精,蔗糖及乳鈣質 ,配製成粉末狀代餐原型。與不含乳清蛋白 生i t制組樣品相較之下,添加實例5中所製備之乳 >月蛋白水解物者之接受度較佳。 實例9 ,签., 每例5呂4^ 的乳清蛋白飲料經調配而含有重量比為8%之於 ,所製備之ALACENTM 392 或 ALACENTM 421 或 ALACE, 〇 y z彳口乳糖夕、β人 酸,香,合物的水解物。該飲料亦含有蔗糖,檸檬 粗& 9 ;及著色劑。該飲料之酸鹼值被調整至4 · 3。該飲 科马一結合5丨 之軟性飲料 蛋白質之營養性及健康性優點且口味清爽 〃' °與含有未處理之乳清蛋白的控制組比較,Page 15 1225490 V. Description of the invention (13) The hydrolysate powder prepared in Example 5 was used, and Gan, Fu Dan and Tian Tiandong were added as yoghurt additives: its: add saccharine to the terminal acid The milk was 2 5% and 5% each. Compared with the control group, the results showed that the lubricity increased and the texture enzymes also changed. Example 7 ° The hydrolysate Bagu from ^… was prepared in Example 5 as early as the Muesli bar recipe, and the water was used as a source of + + 〈poor white shellfish. 6% and 12%. All tasters' preferences for skin% 7 owe to the hydrolysate bar are WPC control group. Among them, the best results were obtained by adding the hydrolyzed ALACENTM 42 1 and ALACENTM ^ She A, which were prepared in Example b. Example 8 = The hydrolysate powder prepared in Example 5 ^ was used as a sample set for the meal replacement concept. The aucentm 42 ™, which is hydrolyzed 7 in ceremonial sugar and protein, is added to the whole milk powder, maltodextrin, sucrose, and milk calcium to make up a powder meal replacement prototype. Compared with the sample made without the whey protein and raw protein, the acceptance of the milk > moon protein hydrolysate prepared in Example 5 was better. Example 9: The amount of each whey protein beverage of 5 to 4 ^ was formulated to contain 8% by weight of the prepared ALACENTM 392 or ALACENTM 421 or ALACE, 〇yz lactose, β-human acid , Fragrant, compound hydrolysate. The drink also contains sucrose, lemon &9; and a coloring agent. The pH of the beverage was adjusted to 4 · 3. This drink Coma combines the nutritional and health benefits of 5 丨 soft drink protein with refreshing taste ° '° compared with the control group containing untreated whey protein,
第16頁 1225490 五、發明說明(14) ' 其對酸驗值之安定性有所改善且該飲料之外觀與牛乳較相 像。 實例1 0 另一營養性的乳清蛋白飲料被調配成含有重量比為 1 2· 5%之於實例5中所製備之ALACENTM 421水解物之水溶 ❿ /夜’在與經巴斯德式殺菌之全乳混合。添加佔最終配方已% 之蔗糖及女定劑。視需要於該飲料中添加相香蕉,香草或 其他類似的香料。得到延長的保存期限,將該飲料經過 1 4 0 C,3秒鐘之超高溫瞬間加熱處理。經過該額外的超高 溫瞬間加熱處理後之平均粒徑仍然為3微米。 實例11 以實例5中之方法進行水解,但以ALACENTM 421粉末還原 液取代新鮮配製之ALACENn ,並游之濃縮成為含3〇%固形、 物之溶液。中性酶之添加量改為〇 . 9 %重量比( 基準),乳糖酶之用量如所界定者。2小時之m貝岸為、昆 合物於固形物15%之下不活化應。ACE—丨活性為〇 48〇公克/ 。其官能性’粒徑及食品應用性均與實例4及實例5極 為近似。 實例1 2 以實例4中之方法進行ALACE評421粉末之水解, : '加率改:9%重量比(以蛋白質為基準)乳糖以得自米 麴囷ASpergl 11US oryzae生產之乳糖酶轉變Page 16 1225490 V. Description of the invention (14) 'The stability of the acid test value has been improved and the appearance of the beverage is similar to milk. Example 10 Another nutrient whey protein beverage was formulated to contain 12.5% by weight of the ALACENTM 421 hydrolysate prepared in Example 5 in a water-soluble solution / night 'in a pasteurized solution. Of whole milk blend. Add sucrose and female fixatives which make up the final formula. Add bananas, vanilla or other similar spices to the beverage as needed. To obtain an extended shelf life, the beverage was subjected to an instantaneous heat treatment at 140 ° C for 3 seconds. The average particle size after this additional ultra-high temperature instantaneous heat treatment was still 3 microns. Example 11 The hydrolysis was carried out in the same manner as in Example 5, but the freshly prepared ALACENn was replaced with ALACENTM 421 powder reducing solution and concentrated to a solution containing 30% solids. The amount of neutral enzyme was changed to 0.9% by weight (baseline), and the amount of lactase was as defined. 2 hours of mbe is that the compound should not be activated below 15% of solids. The activity of ACE- is 0.40 g / g. Its functionality 'particle size and food applicability are very similar to those of Examples 4 and 5. Example 12 The hydrolysis of ALACE 421 powder was carried out in the same manner as in Example 4. The addition rate was changed: 9% by weight (based on protein). Lactose was converted from lactase produced by ASpergl 11US oryzae.
酶30’ 〇〇〇,Kyowa Enzymes Co. Ltd.日本)^ θ A …量比(以乳糖為基準H.5小時之後,該反添=合為 1225490 五、發明說明(15) 物以直接喷入蒸氣使溫度達8 8 °C,維持1. 5或3秒而不活 化。 其粒徑為2 · 3微米。官能性及食品應用性均與實例4之產 物極為近似。 實例1 3 以一商品化酵素:由枯草桿菌生產之中性酶(N 0 v 〇 “『(^31^’丹麥)於酵素濃度為〇.9%重量比時水解含重量比 10%之ALACENTM 392溶液。該反應於50 °C下持續6小時。每 小時取出2 0 0毫升樣品,於8 8 °C下處理8秒以不活化並隨後 將之冷凍乾燥。 ACE- I活性,水解度,溶液之酸鹼值及苦味度隨時間之 發展如下。苦味值愈高則苦味愈強。測定值愈小則ACE_ I 活性愈高。 ^ 表1 : ALACENTM 3 92 WPC 之水解 水解時間 (小時) ACE-Ι活性 (公克/公升) (IC5〇) 水解度 (%) 溶液酸驗值 苦味值 (非正式的, 0-10) 1 0.420 3.86 7.01 0 2 0.280 3.78 6.96 1 3 0.230 4.53 6.92 1 4 0.220 4.89 6.89 3.5 5 0.210 5.20 6.87 2 6 0.190 5.37 6.87 4.5Enzyme 30 ′ 〇〇〇〇, Kyowa Enzymes Co. Ltd. Japan) ^ θ A… the amount ratio (based on lactose H. 5 hours, the reverse addition = 1225490 total 5. Description of the invention (15) to spray directly Into the steam to achieve a temperature of 8 8 ° C, maintaining 1.5 or 3 seconds without activation. Its particle size is 2.3 microns. Functionality and food applicability are very similar to the product of Example 4. Example 1 3 Commercial enzyme: The neutral enzyme (N 0 v 〇 "" (^ 31 ^ 'Denmark) produced by Bacillus subtilis hydrolyzes a solution containing 10% by weight of ALACENTM 392 at an enzyme concentration of 0.9% by weight. This reaction Continue for 6 hours at 50 ° C. Take out 200 ml samples per hour, treat at 8 ° C for 8 seconds to inactivate and freeze freeze dry it later. ACE-I activity, degree of hydrolysis, pH of solution The development of bitterness over time is as follows. The higher the bitterness value, the stronger the bitterness. The smaller the measured value, the higher the ACE_I activity. ^ Table 1: Hydrolysis time (hours) of ALACENTM 3 92 WPC ACE-1 activity (g / Liter) (IC50) Degree of hydrolysis (%) Bitterness value of solution acid (informal, 0-10) 1 0.420 3.86 7.01 0 2 0.280 3.78 6.96 1 3 0.230 4.53 6.92 1 4 0.220 4.89 6.89 3.5 5 0.210 5.20 6.87 2 6 0.190 5.37 6.87 4.5
第18頁 1225490 五、發明說明(16) 實例1 4 以下列商品化蛋白酶於酵素濃度為1 %重量比,5 0 °C下將 含重量比1 0%之ALACENTM 39 2溶液水解1小時。再於88 °C下 處理8秒以不活化並隨後將之冷凍乾燥。 表2: 以不同之酵素水解 酵素 ACE-Ι活性 (公克/公升) (ic5〇) 酸鹼值 水解度 [%] 蛋白酶P6,中性蛋白酶,麴菌屬菌株, 阿米諾酶(Amano Enzymes) 0.274 7.0 8.9 蛋白酶A,中性蛋白酶,米麴菌, 阿米諾酶(Amano Enzymes) 0.443 7.0 9.2 蛋白酶Μ,酸性蛋白酶,米麴議, 阿米諾酶(Amano Enzymes) (L450 4.0 7.4 胜肽酶,中性胜肽酶,米麴菌, 阿米諾酶(Amano Enzymes) 0.540 7.0 6.9 中性酶,中性細菌性蛋白酶,枯草桿菌, Novo Nordisk DK 0.510 7.0 4.3 有效酶(Genancor),酸性真菌性蛋白酶, 黑麴菌,Enzyme Services Ltd. NZ 0.510 4.0 5.6 AFP2000(Genancor),酸性真菌性蛋白 酶,黑麴議,Enzyme Services Ltd. NZ 0.550 4.0 3.9 實例1 5 ACE- I胜肽類之鑑定及其活性測定Page 18 1225490 V. Description of the invention (16) Example 1 4 ALACENTM 39 2 solution containing 10% by weight was hydrolyzed for 1 hour at 50 ° C with an enzyme concentration of 1% by weight at the following commercial protease. It was then treated at 88 ° C for 8 seconds to deactivate and subsequently freeze-dried. Table 2: Hydrolyzing enzyme ACE-1 activity with different enzymes (g / L) (ic50) pH degree of hydrolysis [%] Protease P6, Neutral protease, Pleurotus strain, Amano Enzymes 0.274 7.0 8.9 Protease A, Neutral Protease, Oryzae, Amano Enzymes 0.443 7.0 9.2 Protease M, Acid Protease, Oryza, Amano Enzymes (L450 4.0 7.4 Peptidase , Neutral peptidase, Oryzae, Amano Enzymes 0.540 7.0 6.9 Neutral enzyme, Neutral bacterial protease, Bacillus subtilis, Novo Nordisk DK 0.510 7.0 4.3 Effective enzyme (Genancor), Acid fungal Protease, Black fungus, Enzyme Services Ltd. NZ 0.510 4.0 5.6 AFP2000 (Genancor), acid fungal protease, Heiye, Enzyme Services Ltd. NZ 0.550 4.0 3.9 Example 1 5 Identification and activity of ACE-I peptide Determination
第19頁 1225490 五、發明說明(17) 得自實例3之2 0 0毫克濾過物溶於0. 1%三氟醋酸(TFA)中 並注入一以溶劑A(0· 1%TFA)平衡過之邱比特(Jupiter)製 備性逆相HPLC管柱中(10微米,C18,22x250毫米 [Phenomenex NZ])且連接一 FPLC 系統(Pharmacia)。胜肽 類隨後於0至43%之溶劑B(含0· 08%TFA之乙烯睛)梯度下以 1 0毫升/分鐘之流速於2 4 5分鐘内被沖提出來。沖提液中之 胜〗太類可經由僧測2 1 4 n m之吸光值而得知。沖提液以一自 動化分管收集器於每3分鐘收集一管。Page 19, 1225490 V. Description of the invention (17) 200 mg of the filtrate obtained from Example 3 was dissolved in 0.1% trifluoroacetic acid (TFA) and injected into a solvent A (0.1% TFA) equilibrated. A Jupiter preparative reverse phase HPLC column (10 micron, C18, 22x250 mm [Phenomenex NZ]) was connected to an FPLC system (Pharmacia). The peptides were subsequently flushed out at a flow rate of 10 ml / min in a gradient of 0 to 43% of solvent B (containing 0.08% TFA of vinyl acetate) at a flow rate of 10 ml / min. The victory in the extract can be determined by measuring the absorbance of 2 1 4 n m. The eluate was collected in an automated tube collector every 3 minutes.
將所收集之每一管均加以冷凍乾燥並以重量分析法測定 其所存在之胜肽量。收集物使用體外分析系統(試劑得自Each collected tube was freeze-dried and the amount of peptide present was determined gravimetrically. Collections were performed using an in vitro analysis system (reagents obtained from
Sigma product 30 5- 1 0 )包括兔肺ACE及ACE-Ι活性之分析 乃是利用由白兔比色計量之ACE基質:呋喃丙醯苯基丙氨 醯基甘氨醯基甘胺酸 (Furylacryl〇ylphenyiaianyigiyCylglycine (FApGG)分 析A C E _ 1活性。A C E可將F A P G G 其於340nm處之吸光值下降。 34Onm處之吸光值之變化將會 有最高A C E抑制活性之收集管 管柱中並以較緩梯度之溶劑B 之溶劑B濃度。沖提液以一自 收集一管。 水解得到產物F A P及G G,使得 若一胜肽可抑制ACE,則於 減少。將每毫克胜肽物質具 區分再注入製備性逆相肝^ 沖提’即··每分鐘提高〇. 〇9% 動化分管收集器於每〇· 5分鐘Sigma product 30 5- 1 0) including rabbit lung ACE and ACE-1 activity analysis using the ACE matrix measured by colorimetry of white rabbits: furanyl phenyl alanine glycine glycine glycine (Furylacryl 〇ylphenyiaianyigiyCylglycine (FApGG) analysis of ACE _1 activity. ACE can reduce FAPGG's absorbance at 340nm. The change in absorbance at 34Onm will have the highest ACE inhibitory activity in the collection column and with a slower gradient Concentration of solvent B of solvent B. The eluent is collected by one tube. The products FAP and GG are obtained by hydrolysis, so that if a peptide can inhibit ACE, it is reduced. Each mg of peptide substance is differentiated and then injected into the preparative reverse Xianggan ^ Extraction 'ie.. .9% increase per minute. 9% of the mobilized collector is every 0.5 minutes
將所收集之每一管均以分 並將其中之單一,可鑑別的 區分均加以冷凍乾燥並以重 析式性逆相HPLC管柱予以分析 胜肽收集起來。每一被收集之 量分析法測定其所存在之胜肽Each tube collected was fractionated and a single, identifiable fraction was lyophilized and analyzed on a reversible reverse-phase HPLC column. The peptides were collected. Each peptide collected was used to determine the presence of the peptide
a^2549〇 五、發明說明(18)a ^ 2549〇 5. Description of the invention (18)
里。以前述之方法測定此等經純化之M 並計算個別胜肽之Κ5。。 之胜肽類的ACE-丨活性 每一胜狀之分子量以電子喷霧離子化質譜(^χ Μ 二重四極柱質譜儀)測定之。每—胜狀亦經由μ·之實 ^知描作出Tandem質譜而得到CAD圖譜。每一胜肽亦以自 動化N端定序儀(ABI model 476A蛋白質定序儀)分析。 以此三種技術之所有資料來推測所有具有ace— !活性之胜 肽類。此等具有生物活性之胜肽類來源乃藉由與包含已知 之所有牛乳蛋白序列的資料庫比對而得。 此等胜肽類’其來源’活性及已知之相似度均列於表 3。雖然最後三個胜肽之來源為酪蛋白,但其得自乳清蛋 白之水解物。用以將酪蛋白沉澱之凝乳酶並不使此等酪蛋 白區分沉殿且其仍與乳清蛋白在7一起。in. These purified Ms were determined in the manner described above and the K5 of individual peptides was calculated. . ACE- 丨 activity of the peptides The molecular weight of each peptide was determined by electron spray ionization mass spectrometry (^ χ M double quadrupole mass spectrometer). Each victory also obtained Tandem mass spectrometry through the actual description of μ ·, and obtained CAD map. Each peptide was also analyzed using an automated N-terminal sequencer (ABI model 476A protein sequencer). Based on all the information of these three techniques, we can infer all the peptides with ace-! Activity. These biologically active peptide sources were obtained by alignment with a database containing all known bovine milk protein sequences. The activity of these peptides, their source, and their known similarities are shown in Table 3. Although the source of the last three peptides is casein, it is derived from the hydrolysate of whey protein. The chymosin, which is used to precipitate casein, does not distinguish these caseins from the sink and it is still with whey protein.
第21頁 1225490 五、發明說明(19) 表3. ACE-Ι胜肽類及其活性 胜肽序列a 來源 活性b (K;以毫克公 fh1為單位) 與已知之 ACE-I胜肽之 相似性類 AFE (Ala-Phe-Glu) PPd3(129-131) 20 LFSH (Leu-Phe-Ser-His) PP3(125-128) 30 ILKEKH (Ile-Leu-Lys-Glu-Lys-His) PP3(71-76) 20 LIVTQ (Leu-Ile-Val-Thr-Gln) /3-LGe(l-5) 17 MKG (Met-Lys-Gly) /3-LG(7-9) 24 LDIQKC (Leu-Asp-Ile-Gln-Lys) /3-LG(10-14) 17 /S-LG(9-14) VF (Val-Phe) /3-LG(81-82) 19 ALPMH (Ala-Leu-Pro-Met-His) ;S-LG(142-146) 12 /3-LG(142-148) VTSTAV (Val-Thr-Ser-Thr-Ala-Val) GMPf(59-64) 30 LHLPLP (Leu-His-Leu-Pro-Leu-Pro) /3-CNS(133-138) 7 LVYPFPGPIPNSLPQNIPP (Leu-Val-Tyr-Pro-Phe-Pro-Gly- Pro-Ile-Pro-Asn-Ser-Leu-Pro- Gln-Asn-Ile-Pro-Pro) /?-CN(58-76) 19 /3-CN(74-76) LFRQ (Leu-Phe-Arg-Glu) asrCN(136-139) 17h ❶Page 211225490 5. Description of the invention (19) Table 3. ACE-1 peptides and their active peptide sequences a source activity b (K; in milligrams fh1) is similar to known ACE-I peptides AFE (Ala-Phe-Glu) PPd3 (129-131) 20 LFSH (Leu-Phe-Ser-His) PP3 (125-128) 30 ILKEKH (Ile-Leu-Lys-Glu-Lys-His) PP3 ( 71-76) 20 LIVTQ (Leu-Ile-Val-Thr-Gln) / 3-LGe (l-5) 17 MKG (Met-Lys-Gly) / 3-LG (7-9) 24 LDIQKC (Leu-Asp -Ile-Gln-Lys) / 3-LG (10-14) 17 / S-LG (9-14) VF (Val-Phe) / 3-LG (81-82) 19 ALPMH (Ala-Leu-Pro- Met-His); S-LG (142-146) 12 / 3-LG (142-148) VTSTAV (Val-Thr-Ser-Thr-Ala-Val) GMPf (59-64) 30 LHLPLP (Leu-His- Leu-Pro-Leu-Pro) / 3-CNS (133-138) 7 LVYPFPGPIPNSLPQNIPP (Leu-Val-Tyr-Pro-Phe-Pro-Gly- Pro-Ile-Pro-Asn-Ser-Leu-Pro- Gln- Asn-Ile-Pro-Pro) /?-CN (58-76) 19 / 3-CN (74-76) LFRQ (Leu-Phe-Arg-Glu) asrCN (136-139) 17h ❶
第22頁Page 22
以單字母碼表示 並將相對之三字母碼寫在 胺基酸續列 括狐中 使用有色之基質:fapgg 水解物中含量最多之ACE-I 蛋白酶凍 万一乳球蛋白 _巨胜肽 冷—酪蛋白 人另來源未知之胜肽一起測定活性It is represented by a single letter code and the corresponding three-letter code is written in the amino acid sequel. The colored matrix is used: the ACE-I protease frozen with the most content in fapgg hydrolysate in case of lactoglobulin_Jusheng peptide cold— Casein humans and peptides of unknown origin
實例1 6 =,自發性向血壓大白鼠(shr/n)進行體内血壓試驗以 ,例3中所製備之水解物粉末—(未經超過遽者)之功 放4種大白鼠乃是經過特別之培育使其於成熟期具有高 ^且被廣泛地用來測定降血壓劑之功效。其購自動物 資源中%,p 〇 Box 1180 Canning Vale , Western Australia 6155 。Example 16 = Spontaneous blood pressure test was performed on blood pressure rats (shr / n) spontaneously. The hydrolysate powder prepared in Example 3— (not more than 遽) was used to power amplifier four kinds of rats. It is cultivated to have a high level of maturity and is widely used to determine the efficacy of antihypertensive agents. % Of its purchased animal resources, p 0 Box 1180 Canning Vale, Western Australia 6155.
於試驗中,將八週大之大白鼠個別地關在塑膠鼠籠並置 於有溫控之設備中。無限制地供應其飲水並以商品化之大 白鼠钥料(ad 1 ibi turn)餵養。於八週之飼養期間,每曰以 一定劑量經口餵食試驗產品,並記錄其血壓變化。其血壓 以一特別設計之試驗及血壓測定設備(IITC Inc·,Life Science Instruments, 23924 Victory Blvd, Woodland H i 1 1 d,C A 9 1 3 6 7 )測定之。該實驗之設計乃經梅西In experiments, eight-week-old rats were individually housed in plastic squirrel cages and placed in temperature-controlled equipment. Supply them unlimited water and feed them with a commercial rat ad 1 ibi turn. During the eight-week rearing period, the test product was orally fed at a certain dose every day, and changes in blood pressure were recorded. His blood pressure was measured with a specially designed test and blood pressure measuring device (IITC Inc., Life Science Instruments, 23924 Victory Blvd, Woodland Hi 1 1 d, CA 9 1 3 6 7). The experiment was designed by Messi
(Massey)大學動物倫理 於此八週中,每組動會涊可,案號為98/1 41。 (以最小平方均值表、且物之收縮期血壓變化圖示於圖2中 及4公克/公斤體重^) °飯食水解物2公克/公斤體重/日 只餵食商品化之大白&心兩4組,其收縮期血壓顯著地低於 P<〇.〇〇4,見表3)。雖經最小平方均值分析,其 已知之ACE抑制藥物.;;;!:解物之降血壓功效並不如-/八(蝴去/ ’、 開脫曰(captopril)於施藥量30毫克 a斤胆重/日之效果,<旦對於只餵食商品化之大白鼠飼料 者之血壓已有顯著性的改善。 參考文獻(Massey) University Animal Ethics In each of these eight weeks, each group will be okay, case number 98/1 41. (The table shows the change in systolic blood pressure in the mean square of the least squares, and the graph of the systolic blood pressure changes in Figure 2 and 4 g / kg body weight ^) ° Meals hydrolysate 2 g / kg body weight / day Group, its systolic blood pressure was significantly lower than P < 0.04, see Table 3). Although it is analyzed by means of least squares, its known ACE inhibitory drug. ;;;!: The blood pressure-lowering effect of the solution is not as good as-/ 八 (蝶 去 / ', Captopril) at a dosage of 30 mg a gallbladder The effect of weight / day, < has significantly improved the blood pressure of those who fed only commercial rat feed.
Bernal V & Jelen P (1989), Effectiveness of lactose hydrolysis in Cottage- cheese whey for the development of whey drinks. Mi 1 chwissenchaft 44: 222-225Bernal V & Jelen P (1989), Effectiveness of lactose hydrolysis in Cottage- cheese whey for the development of whey drinks. Mi 1 chwissenchaft 44: 222-225
Cushman D W & Cheung H S(1971),Cushman D W & Cheung H S (1971),
Spectrophotometric assay and properties of the angiotensin converting enzyme in rabbit lung.Spectrophotometric assay and properties of the angiotensin converting enzyme in rabbit lung.
Biochem Pharmacol 20: 1637-1648. FR 2309154, 30 December 1976 Fromageries Bel La •’Biochem Pharmacol 20: 1637-1648. FR 2309154, 30 December 1976 Fromageries Bel La • ’
Vache Qui (From), France. US 3970520, 20 July 1976, General Electric Co, USA. EP0117047, 29 August 1984, General FoodsVache Qui (From), France. US 3970520, 20 July 1976, General Electric Co, USA. EP0117047, 29 August 1984, General Foods
第24頁 1225490 五、發明說明(22)Page 24 1225490 V. Description of the invention (22)
Corporation, USA.Corporation, USA.
Maubois J L, L e on i 1 J, T r ouv e R & Bouha1 1 ab S (19 91) Les peptides du 1 a i t a act i v i t e phys i o1og i que III. Peptides du 1 a i t a effect card i ovascu1 a i re : ac t i v i t e s anti thrombo t i que e t anti hypertensive. La i t, 71, 249 -25 5. JP 4282400, 7 October 1992, Calpis ShokuhinMaubois JL, Le on i 1 J, T rouv e R & Bouha1 1 ab S (19 91) Les peptides du 1 aita act ivite phys i o1og i que III. Peptides du 1 aita effect card i ovascu1 ai re: ac tivites anti thrombo ti que et anti hypertensive. La it, 71, 249 -25 5. JP 4282400, 7 October 1992, Calpis Shokuhin
Kogyο KK, Japan· EP065663, 1 December 1982, Miles LaboratoriesKogyο KK, JapanEP065663, 1 December 1982, Miles Laboratories
Incorporated, USA. JP 8056568, 17 August 1994, Morinaga Milk Co LtdIncorporated, USA. JP 8056568, 17 August 1994, Morinaga Milk Co Ltd
Japan. EP47 45 5 0 6, 1 1 March 1 9 92, Morinaga Milk Co Ltd,Japan. EP47 45 5 0 6, 1 1 March 1 9 92, Morinaga Milk Co Ltd,
Japan. ❿Japan. ❿
Mullally Μ M, Meisel H & FitzGerald R J(1997) Identification of a novel angiotensin-I-converting enzyme inhibitory peptide corresponding to a tryptic fragment of bovine )5 - 1 ac tog 1 obu 1 i n. Federation of European Biochemical Societies Letters, 402, 99-101.Mullally Μ M, Meisel H & FitzGerald RJ (1997) Identification of a novel angiotensin-I-converting enzyme inhibitory peptide corresponding to a tryptic fragment of bovine) 5-1 ac tog 1 obu 1 i n. Federation of European Biochemical Societies Letters , 402, 99-101.
Nakamura Y, Yamamoto N, Sakai K & Takano T ( 1 9 94 ) Antihypertensive effect of the peptides derived from casein by an extracellular proteinase from Lactobacillus helveticus CP7 9 0. Journal of DairyNakamura Y, Yamamoto N, Sakai K & Takano T (1 9 94) Antihypertensive effect of the peptides derived from casein by an extracellular proteinase from Lactobacillus helveticus CP7 9 0. Journal of Dairy
O:\58\58905.ptd 第25頁 1225490 五、發明說明(23)O: \ 58 \ 58905.ptd Page 25 1225490 V. Description of the invention (23)
Science, 77, 917-922.Science, 77, 917-922.
Roy G( 1 9 92 ). Bitterness: reduction and inhibition. Trends in Food Science and Technology 3:85-91Roy G (1 9 92). Bitterness: reduction and inhibition. Trends in Food Science and Technology 3: 85-91
Roy G( 1 997 ). Modifying bitterness: Mechanism, ingredients and applications. Technom i c Publishers, Lancaster, UK. US 4358464, 9 September 1982, Superior DairyRoy G (1 997). Modifying bitterness: Mechanism, ingredients and applications. Technom i c Publishers, Lancaster, UK. US 4358464, 9 September 1982, Superior Dairy
Company, USA.Company, USA.
Yamamoto N ( 1 9 9 7 ). Ant i hyper tensive peptides derived from food proteins. Biopolymers 43: 129-134.Yamamoto N (1 9 9 7). Ant i hyper tensive peptides derived from food proteins. Biopolymers 43: 129-134.
第26頁Page 26
Claims (1)
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
NZ33071098 | 1998-06-17 |
Publications (1)
Publication Number | Publication Date |
---|---|
TWI225490B true TWI225490B (en) | 2004-12-21 |
Family
ID=19926776
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
TW088110663A TWI225490B (en) | 1998-06-17 | 1999-06-24 | Bioactive whey protein hydrolysate |
Country Status (12)
Country | Link |
---|---|
US (4) | US6919314B1 (en) |
EP (1) | EP1087668B1 (en) |
AT (1) | ATE275831T1 (en) |
AU (1) | AU761477B2 (en) |
DE (1) | DE69920219T2 (en) |
DK (1) | DK1087668T3 (en) |
MY (1) | MY126517A (en) |
NO (1) | NO320644B1 (en) |
NZ (1) | NZ508867A (en) |
PE (1) | PE20000717A1 (en) |
TW (1) | TWI225490B (en) |
WO (1) | WO1999065326A1 (en) |
Families Citing this family (45)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1999065326A1 (en) * | 1998-06-17 | 1999-12-23 | New Zealand Dairy Board | Bioactive whey protein hydrolysate |
US6998259B1 (en) | 1999-05-20 | 2006-02-14 | Davisco Foods International | Enzymatic treatment of whey proteins for the production of antihypertensive peptides and the resulting products |
US6630320B1 (en) | 2000-05-08 | 2003-10-07 | Devisco Foods International, Inc. | Treatment of hypertension in mammals with hydrolyzed whey proteins |
NZ506866A (en) * | 2000-09-11 | 2003-05-30 | New Zealand Dairy Board | Bioactive whey protein hydrolysate free of bitter flavours wherein the enzyme used is a heat labile protease |
GB0029777D0 (en) * | 2000-12-06 | 2001-01-17 | Regen Therapeutics Plc | Peptides |
WO2002069734A1 (en) * | 2001-03-05 | 2002-09-12 | Council Of Scientific And Industrial Research | Process for the preparation of protein hydrolysate from milk protein |
FR2827480B1 (en) * | 2001-07-17 | 2003-09-19 | Cie Laitiere Europeenne | MODIFIED WHEY, PREPARATION METHOD, USE AND BREAD PRODUCT COMPRISING MODIFIED WHEY |
WO2003059083A1 (en) * | 2002-01-21 | 2003-07-24 | Fraunhofer-Gesellschaft zur Förderung der angewandten Forschung e.V. | Whey protein hydrolysates and peptide fractions derived from whey protein with a bioactive action, obtained by hydrolysis with proteases |
EP1359157A1 (en) * | 2002-04-29 | 2003-11-05 | Société des Produits Nestlé S.A. | Metallo-proteinase inhibitory agent |
CA2508219C (en) | 2002-12-24 | 2012-12-18 | Her Majesty The Queen In Right Of Canada As Represented By The Minister Of Agriculture And Agri-Food Canada | Ace inhibitory peptides from plant materials |
US7399496B2 (en) * | 2003-02-07 | 2008-07-15 | Glanbia Nutritionals (Ireland) Limited | Hydrolyzed whey protein compositions |
ES2277468B1 (en) * | 2003-05-21 | 2008-07-01 | Grupo Leche Pascual | BIOACTIVE PEPTIDES AND DERIVATIVES, PRODUCTION PROCEDURE, ENTEROCOCCUS FAECALIS STATES PRODUCING SUCH BIOACTIVE PEPTIDES AND ITS APPLICATIONS. |
DE10331202A1 (en) | 2003-07-10 | 2005-03-31 | S.K. Enterprise Gmbh | Use of whey permeate for the treatment of metabolic syndrome |
US20050265946A1 (en) * | 2004-05-28 | 2005-12-01 | Kao Corporation | Elastase inhibitor |
WO2006012536A2 (en) | 2004-07-22 | 2006-02-02 | Ritter Andrew J | Methods and compositions for treating lactose intolerance |
CN100415768C (en) * | 2004-11-29 | 2008-09-03 | 中国农业大学 | Method for producing angiotonin converzyme inhibiting peptide by whey proteinase method |
GB2421890A (en) * | 2004-12-09 | 2006-07-12 | Biocatalysts Ltd | Method for producing a palatability enhancer for a foodstuff |
EP1906769A2 (en) * | 2005-07-15 | 2008-04-09 | Amano Enzyme USA., Ltd. | Enzyme compositions that enhance the flavor of food and beverages |
US20070082056A1 (en) * | 2005-10-12 | 2007-04-12 | Sawhill James W | Antibiotic peptide product |
KR101452756B1 (en) * | 2006-01-04 | 2014-10-22 | 레프리노 푸즈 캄파니 | Protein hydrolysates and method of making |
US7875303B2 (en) * | 2006-03-31 | 2011-01-25 | Kraft Foods Global Brands Llc | Protein system and food products including same |
CN101466389B (en) * | 2006-06-15 | 2013-05-01 | 墨累古尔本合作有限公司 | Formulation comprising whey protein and hydrolysates for improving muscle recovery |
DE102006036285A1 (en) * | 2006-08-03 | 2008-02-07 | "S.U.K." Beteiligungs Gmbh | Whey permeate fractions and their use for the prevention and treatment of type 2 diabetes and metabolic syndrome |
EP1967524A1 (en) * | 2007-03-06 | 2008-09-10 | Friesland Brands B.V. | Methods for producing ACE-inhibitory peptides from whey and peptides obtained thereby |
ES2436145T3 (en) | 2007-11-07 | 2013-12-27 | Mjn U.S. Holdings Llc | Method to reduce bitterness and improve the taste of infant formula without protein and hydrolyzed |
US20110223248A1 (en) * | 2007-12-12 | 2011-09-15 | Ritter Pharmaceuticals, Inc. | Methods and compositions for treating lactose intolerance |
EP2254425A1 (en) | 2008-03-19 | 2010-12-01 | Wisconsin Alumni Research Foundation | Production of protein-polysaccharide conjugates |
WO2009120837A2 (en) | 2008-03-26 | 2009-10-01 | Glanbia Nutritionals (Ireland) Ltd. | Leucine-rich peptide compositions and methods for isolation |
DE102008032828A1 (en) | 2008-07-02 | 2010-01-07 | Technische Universität Dresden | Tryptophan-containing peptides from alpha-lactalbumin with hypotensive and vasoprotective action for biofunctional foods |
US20100055247A1 (en) * | 2008-08-29 | 2010-03-04 | Tirrito Salvatore J | Nutritional Compositions for Athletes |
JP5749419B2 (en) * | 2008-12-24 | 2015-07-15 | 雪印メグミルク株式会社 | Muscle enhancer |
BRPI1008280A2 (en) | 2009-02-24 | 2015-08-25 | Ritter Pharmaceuticals Inc | Prebiotic Formulations and Methods of Use |
NZ595174A (en) * | 2009-03-13 | 2013-04-26 | Univ California | Prebiotic galacto oligosaccharides used to stimulate growth or colonisation of beneficial bifidobacterium bacteria in gut |
CA2773686C (en) | 2009-04-02 | 2018-01-16 | Novozymes A/S | Process for making a milk-based protein hydrolysate |
EP2253324A1 (en) | 2009-04-30 | 2010-11-24 | Consejo Superior De Investigaciones Cientificas | Use of a casein-derived peptide and compositions thereof as antihypertensive |
PT105073A (en) | 2010-04-26 | 2011-10-26 | Consejo Superior Investigacion | PROCESS OF OBTAINING BIOACTIVE PEPTIDIC EXTRACTS THROUGH HYDROLYSIS OF MILK SERUM PROTEINS WITH CYNARA CARDUNCULUS ENZYMES, REFERRING EXTRACTS AND THEIR USES |
EP2563372A4 (en) | 2010-04-28 | 2013-10-02 | Ritter Pharmaceuticals Inc | Prebiotic formulations and methods of use |
FI123201B (en) | 2010-10-29 | 2012-12-14 | Valio Oy | Milk based product and production process |
US20150150275A1 (en) * | 2013-12-03 | 2015-06-04 | The United States Of America, As Represented By The Secretary Of Agriculture | Methods For Processing Acid Whey |
US20190069589A1 (en) | 2015-10-01 | 2019-03-07 | Frieslandcampina Nederland B.V. | Liquid nutritional composition comprising micellar casein and hydrolysed whey protein |
WO2017086303A1 (en) * | 2015-11-16 | 2017-05-26 | キリン株式会社 | Peptide composition and production method therefor |
WO2017102258A1 (en) | 2015-12-18 | 2017-06-22 | Nestec S.A. | Heat sterilized high protein compositions with hydrolyzed protein from a continuous process with at least one endopeptidase |
CN111278293A (en) | 2017-10-26 | 2020-06-12 | 巴斯夫欧洲公司 | Protein hydrolysate as emulsifier for baked food |
JP2022534723A (en) * | 2019-05-29 | 2022-08-03 | アーラ フーズ エエムビエ | Palatable Highly Hydrolyzed Whey Protein Hydrolysate |
CN112010947B (en) * | 2020-09-10 | 2022-05-24 | 中新国际联合研究院 | Cyclohexadecapeptide with angiotensin converting enzyme inhibitory activity and preparation method and application thereof |
Family Cites Families (41)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US592193A (en) * | 1897-10-19 | Sylvania | ||
US3843621A (en) * | 1971-08-19 | 1974-10-22 | Merck & Co Inc | Process for preparing leucyl-leucyl-valyl-henylalanyl methyl ester |
US3970520A (en) | 1973-09-17 | 1976-07-20 | General Foods Corporation | Nutritionally improved foodstuffs |
FR2309154A1 (en) | 1975-03-11 | 1976-11-26 | Bel La Vache Qui Rit Fromage | Recovering protein from lactoserum produced in cheese or casein mfr. - by hydrolysis with beta-galactosidase then ultrafiltration |
US4358464A (en) | 1977-08-02 | 1982-11-09 | Superior Dairy, Inc. | Process for converting sour whey into sweet whey and product |
CH636248A5 (en) * | 1979-03-09 | 1983-05-31 | Nestle Sa | PROCESS FOR THE PREPARATION OF A PURIFIED PROTEIN HYDROLYSATE. |
EP0065663A1 (en) * | 1981-05-11 | 1982-12-01 | Miles Laboratories, Inc. | Method for the preparation of a protein hydrolyzate from whey protein |
US4479975A (en) | 1983-01-13 | 1984-10-30 | General Foods Corporation | Fruit flavored beverages |
US4954443A (en) * | 1984-12-27 | 1990-09-04 | The Nisshin Oil Mills, Ltd. | Method of immobilizing biochemically active substance with xanthan gum |
US4918008A (en) | 1985-01-18 | 1990-04-17 | Gauri Kailash Kumar | Process for the preparation of protein hydrolysate and medicaments containing these hydrolysates |
SE459140B (en) * | 1986-11-25 | 1989-06-12 | Albuglobe Ab | HYDROLYSIS OF WHEAT PROTEIN TO PROVIDE SEPARATE SEPARATION OF FAT |
US4853246A (en) * | 1987-12-01 | 1989-08-01 | Tetra Pak Inc. | Sweetened high protein milk product and process for producing same |
ATE84680T1 (en) | 1987-12-23 | 1993-02-15 | Nestle Sa | PROCESS FOR THE MANUFACTURE OF A WHEY PROTEIN HYDROLYSATE AND A HYPOALLERGENIC FOOD. |
EP0321603A1 (en) | 1987-12-23 | 1989-06-28 | Societe Des Produits Nestle S.A. | Process for preparing a whey protein hydrolyzate and a hypoallergenic foodstuff |
FR2634104B1 (en) | 1988-07-18 | 1991-10-18 | Union Coop Agricole | PARTIAL WHEY PROTEIN HYDROLYSAT, ENZYMATIC PROCESS FOR PREPARING THE SAME, AND HYPOALLERGENIC DIET FOOD CONTAINING THE SAME |
ATE113441T1 (en) | 1989-10-02 | 1994-11-15 | Sandoz Nutrition Ltd | PROTEIN HYDROLYSATES. |
ES2077220T3 (en) | 1990-03-09 | 1995-11-16 | Novo Nordisk As | PROTEIN HYDROLYZES. |
JP2818056B2 (en) | 1990-09-07 | 1998-10-30 | 森永乳業株式会社 | Antimicrobial peptides and antimicrobial agents |
US5093137A (en) * | 1990-10-17 | 1992-03-03 | Shazer Jr William H | Reduced calorie frozen dairy desserts and processes for producing them |
JP3091772B2 (en) * | 1991-03-12 | 2000-09-25 | カルピス株式会社 | Angiotensin converting enzyme inhibiting peptide composition |
JP3149199B2 (en) | 1991-03-12 | 2001-03-26 | カルピス株式会社 | Angiotensin converting enzyme inhibitory peptide |
RU2084172C1 (en) | 1991-05-31 | 1997-07-20 | Данмарк Протеин А/С | Method of preparing the whey protein hydrolyzate |
AU2468292A (en) * | 1991-08-30 | 1993-04-05 | Teagasc, The Agriculture And Food Development Authority | Hypoallergenic whey protein hydrolysate |
JP3092870B2 (en) | 1991-10-18 | 2000-09-25 | 雪印乳業株式会社 | Method for producing milk whey protein hydrolyzate |
DK71292D0 (en) | 1992-05-27 | 1992-05-27 | Novo Nordisk As | |
DE69329923T2 (en) | 1992-11-30 | 2001-09-27 | Morinaga Milk Industry Co. Ltd., Tokio/Tokyo | Whey protein and whey protein hydrolysates, and process for their preparation |
JPH06345664A (en) | 1993-06-11 | 1994-12-20 | Takako Tomita | New composition for suppressing elevation of blood pressure |
US5405637A (en) | 1993-06-30 | 1995-04-11 | Bristol-Myers Squibb Company | Milk protein partial hydrolysate and infant formula containing same |
JP3327500B2 (en) | 1994-08-17 | 2002-09-24 | 森永乳業株式会社 | Lactose-degrading sweet whey concentrate and process for producing the same |
JP3398490B2 (en) * | 1994-09-30 | 2003-04-21 | 雪印乳業株式会社 | Whey protein peptide composition and method for producing the same |
DE69523791T2 (en) | 1994-10-14 | 2002-09-05 | Morinaga Milk Industry Co. Ltd., Tokio/Tokyo | PEPTIDE MIXTURE AND PRODUCTS THEREOF |
JP3542093B2 (en) | 1995-06-01 | 2004-07-14 | オーム乳業株式会社 | Milk composition with low bitterness and low allergen and method for producing the same |
ES2214542T3 (en) * | 1995-06-30 | 2004-09-16 | Arla Foods Amba | METHOD FOR PRODUCING A MIX OF PEPTIDES. |
JPH1033115A (en) | 1996-07-24 | 1998-02-10 | Nouchikusangiyou Shinko Jigyodan | Whey beverage and its production |
US5837312A (en) | 1996-09-20 | 1998-11-17 | Abbott Laboratories | Debittered protein product having improved antigenicity and method for manufacture |
EP0996857B1 (en) | 1997-07-17 | 2009-09-09 | Ludwig Institute For Cancer Research | Cancer associated nucleic acids and polypeptides |
WO1999065326A1 (en) * | 1998-06-17 | 1999-12-23 | New Zealand Dairy Board | Bioactive whey protein hydrolysate |
MY129566A (en) | 1999-01-19 | 2007-04-30 | Nestle Sa | A hypoallergenic composition containing tolerogenic peptides inducing oral tolerance |
US6630320B1 (en) * | 2000-05-08 | 2003-10-07 | Devisco Foods International, Inc. | Treatment of hypertension in mammals with hydrolyzed whey proteins |
US6551636B2 (en) * | 1999-07-23 | 2003-04-22 | Novozymes A/S | Modification of foaming properties of proteins |
CA2415688A1 (en) | 2000-05-08 | 2001-11-15 | Anand Rao | Enzymatic treatment of whey proteins for the production of antihypertensive peptides, the resulting products and treatment of hypertension in mammals |
-
1999
- 1999-06-14 WO PCT/NZ1999/000084 patent/WO1999065326A1/en active IP Right Grant
- 1999-06-14 EP EP99928257A patent/EP1087668B1/en not_active Expired - Lifetime
- 1999-06-14 AU AU45359/99A patent/AU761477B2/en not_active Ceased
- 1999-06-14 AT AT99928257T patent/ATE275831T1/en not_active IP Right Cessation
- 1999-06-14 DE DE69920219T patent/DE69920219T2/en not_active Expired - Lifetime
- 1999-06-14 NZ NZ508867A patent/NZ508867A/en unknown
- 1999-06-14 DK DK99928257T patent/DK1087668T3/en active
- 1999-06-14 US US09/720,041 patent/US6919314B1/en not_active Expired - Fee Related
- 1999-06-16 MY MYPI99002487A patent/MY126517A/en unknown
- 1999-06-16 PE PE1999000532A patent/PE20000717A1/en not_active Application Discontinuation
- 1999-06-24 TW TW088110663A patent/TWI225490B/en not_active IP Right Cessation
-
2000
- 2000-12-12 NO NO20006305A patent/NO320644B1/en unknown
-
2001
- 2001-09-11 US US10/070,406 patent/US7148034B2/en not_active Expired - Fee Related
-
2005
- 2005-03-17 US US11/083,662 patent/US20050164340A1/en not_active Abandoned
-
2009
- 2009-02-23 US US12/391,220 patent/US20090258387A1/en not_active Abandoned
Also Published As
Publication number | Publication date |
---|---|
WO1999065326A1 (en) | 1999-12-23 |
DE69920219D1 (en) | 2004-10-21 |
DK1087668T3 (en) | 2005-01-24 |
EP1087668B1 (en) | 2004-09-15 |
NO20006305L (en) | 2001-02-14 |
US7148034B2 (en) | 2006-12-12 |
DE69920219T2 (en) | 2005-09-22 |
US20090258387A1 (en) | 2009-10-15 |
US6919314B1 (en) | 2005-07-19 |
AU761477B2 (en) | 2003-06-05 |
PE20000717A1 (en) | 2000-08-04 |
ATE275831T1 (en) | 2004-10-15 |
NO20006305D0 (en) | 2000-12-12 |
EP1087668A1 (en) | 2001-04-04 |
AU4535999A (en) | 2000-01-05 |
NO320644B1 (en) | 2006-01-09 |
NZ508867A (en) | 2003-11-28 |
US20040086958A1 (en) | 2004-05-06 |
US20050164340A1 (en) | 2005-07-28 |
MY126517A (en) | 2006-10-31 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
TWI225490B (en) | Bioactive whey protein hydrolysate | |
EP1317186B1 (en) | Improved bioactive whey protein hydrolysate | |
Abd El-Salam et al. | Preparation, properties, and uses of enzymatic milk protein hydrolysates | |
RU2292155C2 (en) | Method for milk proteins hydrolysis | |
AU2001290376A1 (en) | Improved bioactive whey protein hydrolysate | |
JP5924653B2 (en) | Heat-pasteurized or heat-sterilized packaged composition | |
US20140179609A1 (en) | Whey protein hydrolysate containing tryptophan peptide consisting of alpha lactalbumin and the use thereof | |
JP5695326B2 (en) | Protein synthesis promoter | |
JP2959747B2 (en) | Savory whey protein hydrolyzate and method for producing the same | |
Perotti et al. | Biomolecules Derived from Whey: Strategies for Production and Biological Properties | |
JPH0678685A (en) | Production of protein hydrolyzate without unpleasant taste | |
AU2003204321B2 (en) | Bioactive Whey Protein Hydrolysate | |
BG62507B1 (en) | Low-lactose dietetic dairy product and method for its preparation | |
JP2003339326A (en) | Whey protein hydrolyzate and method for producing the same | |
Khan | Received: 19 th Jan-2013 Revised: 31 st Jan-2013 Accepted: 2 nd Feb-2013 Research article WHEY PROTEIN HYDROLYSATES: TECHNO-FUNCTIONAL PERSPECTIVE |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
MM4A | Annulment or lapse of patent due to non-payment of fees |